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Journal of Clinical Microbiology, June 2000, p. 2200-2203, Vol. 38, No. 6
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

rpoB Gene Analysis as a Novel Strategy for Identification of Spirochetes from the Genera Borrelia, Treponema, and Leptospira

Patricia Renesto, Katell Lorvellec-Guillon, Michel Drancourt, and Didier Raoult*

Unité des Rickettsies, CNRS UPRES-A 6020, Faculté de Médecine, Université de la Méditerranée, Marseille, France

Received 27 January 2000/Returned for modification 11 March 2000/Accepted 31 March 2000

Spirochetes are emerging pathogens for which culture and identification are partly unresolved. In fact, 16S rRNA-based sequencing is by far the most widely used PCR methodology that is able to detect such uncultivable pathogens. However, this assay actually has some limitations linked to potential problems of contamination, which hampers diagnosis. To circumvent this, we have devised a simple PCR strategy involving targeting of the gene encoding the RNA polymerase beta subunit (rpoB), a highly conserved enzyme. The complete sequence of the Leptospira biflexa (serovar patoc) rpoB gene was determined and compared with the published sequences for Borrelia burgdorferi and Treponema pallidum. From the resulting analysis, degenerate nucleotide primers were designed and tested for their ability to amplify a portion of the rpoB gene from various spirochetes. Using two different pairs of these primers, we succeeded in obtaining specific rpoB-amplified fragments for all members of the genera Leptospira, Treponema, and Borrelia tested and no other bacteria. Our findings may have significant implications for the development of a new tool for the identification of spirochetes, especially if clinical samples are contaminated or when the infecting strain is uncultivable.

* Corresponding author. Mailing address: Unité des Rickettsies, CNRS UPRES-A 6020, Faculté de Médecine, Université de la Mediterranée, Marseille, France. Phone: 33 4 91 83 43 75. Fax: 33 4 91 83 03 90. E-mail: Didier.RAOULT{at}medecine.univ-mrs.fr.

Journal of Clinical Microbiology, June 2000, p. 2200-2203, Vol. 38, No. 6
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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